Transcriptional Profiling and Dynamical Regulation Analysis Identify Potential Kernel Target Genes of SCYL1-BP1 in HEK293T Cells

SCYL1-BP1 is thought to function in the p53 pathway through Mdm2 and hPirh2, and mutations in SCYL1-BP1 are associated with premature aging syndromes such as Geroderma Osteodysplasticum; however, these mechanisms are unclear. Here, we report significant alterations in miRNA expression levels when SCYL1-BP1 expression was inhibited by RNA interference in HEK293T cells. We functionally characterized the effects of potential kernel miRNA-target genes by miRNA-target network and protein-protein interaction network analysis. Importantly, we showed the diminished SCYL1-BP1 dramatically reduced the expression levels of EEA1, BMPR2 and BRCA2 in HEK293T cells. Thus, we infer that SCYL1-BP1 plays a critical function in HEK293T cell development and directly regulates miRNA-target genes, including, but not limited to, EEA1, BMPR2, and BRCA2, suggesting a new strategy for investigating the molecular mechanism of SCYL1-BP1.     

Effects of Multi-nutrient Additions on GHG Fluxes in a Temperate Grassland of Northern China

Human activities have substantially enhanced the availability of important nutrient elements such as nitrogen (N), phosphorus (P), and potassium (K) in ecosystems worldwide. However, how the concurrent increase in all of these nutrients will affect greenhouse gas (that is, CO₂, N₂O, CH₄) levels remains unknown. In a temperate steppe of northern China, a 2-year field experiment was conducted to examine the effects of multi-nutrient additions on GHG fluxes from 2009 to 2010. Four levels of annual nutrient loads were mimicked: 0 g NPK (control), 15.5 g P m^?2 and 19.5 g K m^?2 as KH₂PO₄ (PK), 10 g N m^?2 as NH₄NO₃ plus PK (10N + PK), and 20 g N m^?2 plus PK (20N + PK) per year. The results show that multi-nutrient additions led to significant increases in net primary production (NPP) and soil temperature (ST), a significant decrease in soil moisture (SM) in 2010, and no significant changes in other soil parameters. Seasonal patterns differed greatly for different GHG fluxes in response to different nutrient treatments, largely as a result of differences in influential factors. The 10N + PK treatment significantly increased CO₂ uptake, whereas the 20N + PK treatment significantly decreased CO₂ uptake. The application of P and K without additional N significantly enhanced CH₄ uptake, whereas the two N + PK treatments significantly enhanced N₂O emissions. Significant positive, linear relationships were found between cumulative CO₂ uptake and soil total nitrogen (TN), microbial biomass carbon, and microbial biomass nitrogen, whereas significant negative, linear relationships were found with NPP, SM, and the C/N ratio. Significant positive, linear relationships were found between cumulative N₂O emission and ST, TN, NPP, and total organic carbon, whereas no relationships were found between cumulative CH₄ uptake and any soil parameters. CO₂ flux was related to N₂O flux temporally, to a certain extent, for all the treatments. In the control, N₂O flux showed a negative, linear relationship with CH₄ flux, whereas no regular relationships were detected between CO₂ and CH₄ fluxes in any treatment. Our findings imply that increasing nutrient deposition will change the magnitude, patterns, and relationships among GHG uptakes and emissions in the future.     

The Ubiquitination of Spinal MrgC Alleviates Bone Cancer Pain and Reduces Intracellular Calcium Concentration in Spinal Neurons in Mice

Neurochemical Research - Mas-related G-protein-coupled receptor subtype C (MrgC) has been shown to play an important role in the development of bone cancer pain. Ubiquitination is reported to...     

Generation and Efficacy Evaluation of Recombinant Classical Swine Fever Virus E2 Glycoprotein Expressed in Stable Transgenic Mammalian Cell Line

Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), which is a highly contagious swine disease that causes significant economic loses to the pig industry worldwide. The envelope E2 glycoprotein of CSFV is the most important viral antigen in inducing protective immune response against CSF. In this study, we generated a mammalian cell clone (BCSFV-E2) that could stably produce a secreted form of CSFV E2 protein (mE2). The mE2 protein was shown to be N-linked glycosylated and formed a homodimer. The vaccine efficacy of mE2 was evaluated by immunizing pigs. Twenty-five 6-week-old Landrace piglets were randomly divided into five groups. Four groups were intramuscularly immunized with mE2 emulsified in different adjuvants twice at four-week intervals. One group was used as the control group. All mE2-vaccinated pigs developed CSFV-neutralizing antibodies two weeks after the first vaccination with neutralizing antibody titers ranging from 1∶40 to 1∶320. Two weeks after the booster vaccination, the neutralizing antibody titers increased greatly and ranged from 1∶10,240 to 1∶81,920. At 28 weeks after the booster vaccine was administered, the neutralizing antibody titers ranged from 1∶80 to 1∶10240. At 32 weeks after the first vaccination, pigs in all the groups were challenged with a virulent CSFV strain at a dose of 1×10⁵ TCID₅₀. At two weeks after the challenge, all the mE2-immunized pigs survived and exhibited no obvious symptoms of CSF. The neutralizing antibody titer at this time was 20,480. Unvaccinated pigs in the control group exhibited symptoms of CSF 3–4 days after challenge and were euthanized from 7–9 days after challenge when the pigs became moribund. These results indicate that the mE2 is a good candidate for the development of a safe and effective CSFV subunit vaccine.